Fusion polynucleotide for biosynthesis of beta-carotene using bicistronic gene expression and method for producing beta-carotene using the same

Abstract

The present invention relates to fusion polynucleotides for biosynthesis of beta-carotene and a method for producing beta-carotene using the same. More particularly, it relates to fusion polynucleotides encoding phytoene synthase, connective sequences of FMDV-derived 2A sequence or internal ribosome entry site (IRES) and carotene desaturase, and a method for producing beta-carotenes using the same. Fusion polynucleotides and the recombinant vector using the same of the present invention have the effect of expression both phytoene synthase gene and carotene desaturase gene stably within cell transformants. Accordingly, fusion polynucleotides of the present invention can be used to regulate the biosynthetic metabolism of plant producing beta-carotene. Furthermore, it can be applied to effectively increase the content of beta-carotene, a useful metabolite.

The invention claimed is: 
     
       1. A recombinant vector comprising a promoter and a fusion polynucleotide for the biosynthesis of beta-carotene comprising the nucleotide sequence as set forth in SEQ ID NO:1 having the phytoene synthase gene of pepper, FMDV-derived 2A sequence optimized in rice plant, and the carotene desaturase gene of bacteria, wherein the recombinant vector is a pGIb-PIC vector or a pGIb-PAC vector. 
     
     
       2. A recombinant vector comprising a promoter and a fusion polynucleotide for the biosynthesis of beta-carotene comprising the nucleotide sequence as set forth in SEQ ID NO:42 having the phytoene synthase gene of pepper, the internal ribosome entry site (IRES) gene, and the carotene desaturase gene of bacteria, wherein the recombinant vector is a pGIb-PIC vector or a pGIb-PAC vector. 
     
     
       3. A transformed cell, which is transformed with a recombinant vector selected from the group consisting of the recombinant vectors according to  claim 1  and  claim 2 . 
     
     
       4. The transformed cell of  claim 3 , which is  Agrobacterium tumefaciens  or  Agrobacterium rhizogene.    
     
     
       5. A transformed plant cell, which is transformed with a vector selected from the group consisting of the recombinant vectors according to  claim 1  and  claim 2  to synthesize β-carotene. 
     
     
       6. A transformed plant, which is transformed with a vector selected from the group consisting of the recombinant vectors according to  claim 1  and  claim 2  to synthesize β-carotene. 
     
     
       7. A plant of  claim 6 , which is mono-cotyledonous or di-cotyledonous. 
     
     
       8. A plant of  claim 6 , wherein the plant is selected from the group consisting of rice plant, wheat, barley, bamboo shoot, corn, taro, asparagus, onion, garlic, welsh onion, scallion, wild rocambole, yam, ginger,  Arabidopsis , eggplant, tobacco, pepper, tomato, burdock, crown daisy, lettuce, bellflower, spinach, spinach beet, sweet potato, celery, carrot, dropwort, parsley, white cabbage, cabbage, radish, water melon, melon, cucumber, pumpkin, gourd, strawberry, soy bean, mung bean, kidney bean, bird's-foot trefoil, potato, duckweed, green perilla, pigeon pea, narcissus, marigold and green bean. 
     
     
       9. A transformed mushroom, which is transformed with a vector selected from the group consisting of the recombinant vectors according to  claim 1  and  claim 2  to synthesize β-carotene.